The purpose of this study was to evaluate nurses’ knowledge of infection prevention procedures, the degree to which they were applied correctly, and whether length of service affected either knowledge or application. Nurses with over five years of experience had significantly increased understanding of infection prevention (p=0.009) and significantly increased application of knowledge to practice (p=0.001), compared to nurses with five years or less experience. In particular, understanding of hand hygiene and use of personal protective equipment (PPE) was poor although application was compliant, while knowledge of care of patients with meticillin resistant Staphylococcus aureus (MRSA) and Clostridium difficile was poor, which was reflected by substandard application of knowledge to practice. The results of this study suggest that focusing infection prevention education around patients with specific infections, such as MRSA and C. difficile, rather than on individual standard precautions may more effectively increase knowledge and therefore application of infection prevention practices